Determination of intracellular esterase activity using ratiometric Raman sensing and spectral phasor analysis

Braddick, H. J., Tipping, W., Wilson, L. T. , Jaconelli, H., Grant, E. K., Faulds, K., Graham, D. and Tomkinson, N. (2023) Determination of intracellular esterase activity using ratiometric Raman sensing and spectral phasor analysis. Analytical Chemistry, 95(12), pp. 5369-5376. (doi: 10.1021/acs.analchem.2c05708) (PMID:3692685) (PMCID:PMC10061367)

Abstract

Carboxylesterases (CEs) are a class of enzymes that catalyze the hydrolysis of esters in a variety of endogenous and exogenous molecules. CEs play an important role in drug metabolism, in the onset and progression of disease, and can be harnessed for prodrug activation strategies. As such, the regulation of CEs is an important clinical and pharmaceutical consideration. Here, we report the first ratiometric sensor for CE activity using Raman spectroscopy based on a bisarylbutadiyne scaffold. The sensor was shown to be highly sensitive and specific for CE detection and had low cellular cytotoxicity. In hepatocyte cells, the ratiometric detection of esterase activity was possible, and the result was validated by multimodal imaging with standard viability stains used for fluorescence microscopy within the same cell population. In addition, we show that the detection of localized ultraviolet damage in a mixed cell population was possible using stimulated Raman scattering microscopy coupled with spectral phasor analysis. This sensor demonstrates the practical advantages of low molecular weight sensors that are detected using ratiometric Raman imaging and will have applications in drug discovery and biomedical research.

• Item Type: Articles
• Additional Information: We thank the University of Strathclyde, the EPSRC (EP/TR512114/1; EP/V519777/1), and GlaxoSmithKline for financial support.
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Wilson, Mr Liam
• Authors: Braddick, H. J., Tipping, W., Wilson, L. T., Jaconelli, H., Grant, E. K., Faulds, K., Graham, D., and Tomkinson, N.
• College/School: College of Science and Engineering > School of Chemistry
• Journal Name: Analytical Chemistry
• Publisher: American Chemical Society (ACS)
• ISSN: 0003-2700
• ISSN (Online): 1520-6882
• Published Online: 16 March 2023
• Copyright Holders: Copyright © 2023 The Authors
• First Published: First published in Analytical Chemistry 95(12):5369-5376
• Publisher Policy: Reproduced under a Creative Commons license